Cardiac alpha-crystallin. III. Involvement during heart ischemia.
Rat hearts were perfused in the working heart or Langendorff mode and then subjected to total normothermic ischemia. The content of alpha-crystallin in the water soluble protein fraction obtained from these hearts diminished in a time-dependent manner during ischemia. The protein was recovered in the low g pellet of the homogenate. The redistribution was dramatic, selective for alpha-crystallin and irreversible. Large crystallin clumps formed also when exposing the soluble protein fraction of control hearts to slightly acidic pH (6.5-7.0). Electron microscopic analysis showed that aggregation of the globular homo-oligomeric units of crystallin occurred. The aggregates probably represented denatured protein and were similar in appearance to lenticular alpha H-crystallin. In purified form, however, cardiac crystallin particles did not cluster at pH 6.5. Aggregation only occurred in the presence of other protein components (including, probably, cytosolic actin) of the soluble fraction. A direct and selective interaction between actin and cardiac crystallin could be demonstrated using actin-Sepharose affinity chromatography procedures. The results suggest that large aggregates of cardiac crystallin form very early during ischemia, due to acidification of the cytosol. Cardiac crystallin is highly homologous to stress proteins and is localized on the Z-disks, where it plays probably a structural or protective role. Its rapid and complete denaturation could be involved in the genesis of the irreversible structural damages occurring during ischemia.
Chiesi
M
M
Department of Research, Pharmaceuticals Division, Ciba-Geigy Ltd, Basel, Switzerland.
Longoni
S
S
Limbruno
U
U
D000818:Animals; D003327:Coronary Disease / Q000378:metabolism* / Q000473:pathology; D003459:Crystallins / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism / Q000648:ultrastructure; D009119:Muscle Contraction; D011489:Protein Denaturation; D051381:Rats; D011919:Rats, Inbred Strains; D012995:Solubility
D003459:Crystallins